Studies of hepatitis C virus polarized cell entry and host factor requirements

丙型肝炎病毒极化细胞进入和宿主因子要求的研究

• 批准号: 7448969
• 负责人: Matthew J Evans
• 金额: $ 9万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-15 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7448969
• 关键词: Adverse effects; Animal Model; Antiviral Agents; Antiviral Therapy; Binding; CD81 gene; Caco-2 Cells; Cell Line; Cell Polarity; Cells; Clathrin; Complex; Cultured Cells; Development; Effectiveness; Endogenous Factors; Event; Exhibits; Fibrosis; Foundations; Future; Genetic; Genotype; Glycoproteins; Glycosaminoglycans; Goals; HCV screening; Head; Hepatitis C; Hepatitis C virus; Hepatocyte; Human; Individual; Infection; Integration Host Factors; Interferons; Intestines; Investigation; Label; Laboratory Research; Life Cycle Stages; Liver Failure; Liver diseases; Location; Mediating; Mus; Mutation; Nature; Phase; Play; Process; Proteins; Public Health; RNA replication; Recycling; Research; Research Personnel; Retroviridae; Role; SR-BI receptor; System; Temperature; Tight Junctions; Transplantation; United States; Viral; Viral Proteins; Virion; Virus; Virus Diseases; Virus Replication; Work; anti-hepatitis C; base; career; cell type; claudin-1 protein; env Gene Products; expression cloning; graft failure; human PHEMX protein; improved; inhibitor/antagonist; innovation; insight; liver transplantation; monolayer; mutant; novel; particle; pathogen; polarized cell; prevent; programs; receptor; research study

DESCRIPTION (provided by applicant): Hepatitis C virus (HCV) is the leading cause of liver disease in the United States. With no specific anti-HCV therapies, the currently employed interferon-based treatment is inadequate, as it has severe side effects and is only effective in half of the major genotype infected individuals. My past, current, and future research is aimed at understanding the HCV replication mechanisms, with the ultimate goal of uncovering novel antiviral targets. In the past, I have studied how HCV replication is regulated, how viral proteins interact with each other and cellular factors, and helped establish systems to study HCV in cell culture. Currently, my research is focused on the earliest events in the viral life cycle involving entry into the host cell, which are poorly understood processes. In particular, although several host factors have been implicated as involved in HCV entry, little is know as to how they are utilized by the virus. Our recent identification of the tight junction protein, claudin-1 (CLDN1), as essential for this process was a major development in this field. This discovery has opened-up a whole new view of HCV cell entry, as the involvement of CLDN1 in cell polarity strongly suggests that the polarized nature of an hepatocyte may influence how HCV enter a cell. This proposal describes experiments to: 1) study HCV entry into polarized cells, 2) define how the virion interacts with and utilizes the known HCV entry factors, and 3) perform additional screens for HCV entry factors able to render both human and murine cells infectable with HCV. Such investigations will provide greater, much needed insight in HCV replication, as well as lay the foundation for future studies of replication of HCV and other related viruses, which I will pursue further in the independent phase of my career as the head of an academic research laboratory. Liver failure from HCV is the leading cause of liver transplantation, which is often unsuccessful due to universal reinfection after transplantation, frequently resulting in rapid fibrosis progression and subsequent graft failure. The proposed experiments, aimed at further understanding the HCV entry process, are directly related to the development of novel antiviral therapies to inhibit HCV cell entry, which could prevent graft reinfection and thus greatly improve the effectiveness of liver transplantation. In addition, this work is directly related to the development of much needed HCV small animal models.

描述（由申请人提供）：丙型肝炎病毒（HCV）是美国肝脏疾病的主要原因。由于没有特异性的抗HCV疗法，目前采用的基于干扰素的治疗是不够的，因为它具有严重的副作用，并且仅对一半的主要基因型感染个体有效。我过去、现在和未来的研究旨在了解丙型肝炎病毒复制机制，最终目标是发现新的抗病毒靶点。过去，我研究了 HCV 复制是如何调节的、病毒蛋白之间以及细胞因子之间如何相互作用，并帮助建立了在细胞培养中研究 HCV 的系统。目前，我的研究重点是病毒生命周期中涉及进入宿主细胞的最早事件，这是人们知之甚少的过程。特别是，尽管一些宿主因素被认为与丙型肝炎病毒的进入有关，但人们对病毒如何利用它们知之甚少。我们最近鉴定出紧密连接蛋白 Claudin-1 (CLDN1) 对于这一过程至关重要，这是该领域的一项重大进展。这一发现开辟了 HCV 细胞进入的全新视角，因为 CLDN1 参与细胞极性强烈表明肝细胞的极化性质可能影响 HCV 进入细胞的方式。该提案描述了以下实验：1) 研究 HCV 进入极化细胞，2) 定义病毒粒子如何与已知的 HCV 进入因子相互作用并利用已知的 HCV 进入因子，以及 3) 对能够使人类和小鼠细胞都感染的 HCV 进入因子进行额外的筛选与丙肝病毒。此类研究将为 HCV 复制提供更广泛、急需的见解，并为未来 HCV 和其他相关病毒复制的研究奠定基础，我将在作为学术研究负责人的职业生涯的独立阶段进一步追求这些研究实验室。 HCV 引起的肝衰竭是肝移植的主要原因，肝移植常常由于移植后普遍的再感染而失败，常常导致快速纤维化进展和随后的移植失败。所提出的实验旨在进一步了解HCV进入过程，与抑制HCV细胞进入的新型抗病毒疗法的开发直接相关，这可以防止移植物再感染，从而大大提高肝移植的有效性。此外，这项工作与急需的HCV小动物模型的开发直接相关。